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For example, there are many cases in which cultured murine and human EC cells are used to study the process of neural differentiation [Andrews, 1984; Jones-Villeneuve et al, 1982; Macpherson et al, 1995; McBurney et al, 1988; Przyborski et al, 2000; Stewart et al, 2003; see also 3] The more restricted developmental potential of certain EC cell lines can offer an alternative to primary cells, where consistency and quantity of material are variable, and embryonic stem cells, which are technically more challenging to grow and have the tendency to spontaneously differentiate and form progeny representative of all three germ layers Research using human EC cells as models of early embryogenesis remains a useful, viable alternative and serves as a simple and robust experimental system to investigate, for example, cell fate determination in the embryonic ectoderm [see for review Przyborski et al, 2004] 61.

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2 Culture of Human Embryonal Carcinoma Cell Lines In general, it is possible to isolate and establish EC cell lines from teratocarcinomas whatever their origin providing there is a resident population of proliferating stem cells Another generalization is the morphology of such cells, in that all murine and human EC lineages derived so far share a characteristic structure of a sparse ring of cytoplasm surrounding a rounded nucleus with one or two prominent nucleoli Moreover, such morphology closely resembles that of embryonic stem cells EC cells generally grow as tightly packed colonies, with cells adhering strongly to one another, and often the boundaries between cells are indistinct Many of the widely used EC cell lines have adapted to grow independent of feeder cells Thus it is technically less challenging to grow EC cells routinely, and this allows the generation of large quantities of homogeneous cellular material.

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In general, the requirements for culture of human EC cells are modest compared to human embryonic stem cells, in that specialized (and often expensive) media supplements and feeder cells are not needed Dulbecco s modi ed Eagle s medium (high-glucose formulation supplemented with 2 mM l-glutamine) is most commonly used as a base medium to grow EC cells, although other investigators have preferred the -modi cation of Eagle s minimal essential medium ( -MEM) The medium is usually supplemented by 10% heat-inactivated fetal bovine serum (FBS) It is recommended that different batches of FBS from alternative suppliers are screened to select a batch that is optimal for growth and differentiation by examining several for EC cell growth and ability to differentiate before selection of the batch.

Notice that Excel displays the data series as concentric rings. As you can see, a doughnut chart with more than one series can be very difficult to interpret. For example, the relatively larger sizes of the slices toward the outer part of the doughnut can be deceiving. Consequently, you should use doughnut charts sparingly. Perhaps the best use for a doughnut chart is to plot a single series as a visual alternative to a pie chart. In many cases, a stacked column chart for such comparisons expresses your meaning better than does a doughnut chart (see Figure 19.28).

We look at the expression of cell surface stem cell markers (SSEA-3, TRA1-60) and differentiation markers (VINIS, A2B5, after 7-day retinoic acid exposure) by ow cytometry (see Protocol 65) Standard tissue culture plastic asks and multiwelled plates from any of the major suppliers of consumables are generally satisfactory for.

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[chieh@penguin home]$ dir total 132 drwxr-xr-x 2 4096 Mar 11 00:28 ./ drwxr-xr-x 20 4096 Mar 11 00:28 ../ -rw-r--r-1 122687 Mar 10 23:58 photopc-3.05.tar.gz [chieh@penguin home]$ tar xvzf photopc-3.05.tar.gz

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6. Click the OK button to dismiss the Conditional Formatting dialog. 7. With the asterisk text object still selected, choose Format Text Color and select a light gray to match the color of the tab panel behind the asterisk.

where n and p denote neutrons and protons, respectively. Neutral-current processes may also be studied, but we do not reproduce the reIevant formulas.

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